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目的研究Hedgehog(Hh)信号通路阻断剂环王巴明对脑胶质瘤U87细胞增殖及凋亡的影响。方法体外培养U87细胞,以10、50、100μmol/L的环王巴明作用24h,MTT法检测环王巴明对U87细胞增殖的影响,流式细胞技术检测环王巴明对U87细胞凋亡的影响,RT-PCR检测环王巴明对U87细胞Gli-1 mRNA表达的变化,Western blot检测环王巴明对U87细胞Gli-1蛋白表达的变化。结果 MTT检测结果表明10μmol/L环王巴明组(85.11±2.61)%、50μmol/L环王巴明组(76.81±3.13)%和100μmol/L环王巴明组(69.20±4.11)%细胞活性较空白对照组(99.87±0.21)%降低(P<0.05);流氏细胞技术检测结果表明10μmol/L环王巴明组(15.22±0.21)%、50μmol/L环王巴明组(24.87±0.24)%和100μmol/L环王巴明组(31.36±0.26)%细胞总凋亡率较空白对照组(2.66±0.28)%升高(P<0.05);RT-PCR检测结果表明10μmol/L环王巴明组(0.68±0.06)%、50μmol/L环王巴明组(0.48±0.06)%和100μmol/L环王巴明组(0.30±0.06)%Gli-1mRNA表达水平较空白对照组(0.92±0.11)%降低(P<0.05);Western blot检测结果表明10μmol/L环王巴明组(0.78±0.06)%、50μmol/L环王巴明组(0.58±0.05)%和100μmol/L环王巴明组(0.40±0.04)%Gli-1蛋白表达水平较空白对照组(0.94±0.11)%降低(P<0.05)。结论Hedgehog信号通路阻断剂环王巴明可以抑制脑胶质瘤U87细胞增殖、诱导U87细胞凋亡,其作用机制可能与下调GLi-1表达有关。
Objective To study the effects of Hedgehog (Hh) signaling pathway inhibitor Wangbaming on proliferation and apoptosis of glioma U87 cells. Methods U87 cells were cultured in vitro, and treated with 10, 50 and 100 μmol / L of eupramine for 24 h. The proliferation of U87 cells was detected by MTT assay. The apoptosis of U87 cells was detected by flow cytometry The changes of Gli-1 mRNA expression in U87 cells were detected by RT-PCR. The expression of Gli-1 protein in U87 cells was detected by Western blot. Results The results of MTT assay showed that the cells in the group of 10 μmol / L Wangbaming (85.11 ± 2.61)%, 50 μmol / L (26.81 ± 3.13)% and 100 μmol / L (69.20 ± 4.11)% (99.87 ± 0.21)% (P <0.05). The results of flow cytometry showed that the inhibitory effect of 10 μmol / L cyclopentamine (15.22 ± 0.21)% and 50 μmol / L cyclopentamine (24.87% (P <0.05). The results of RT-PCR showed that the apoptosis rate of 10μmol / L Cyclobalan group (31.36 ± 0.26)% was significantly higher than that of blank control group (2.66 ± 0.28% The expression of Gli-1mRNA in L-eupamine group (0.68 ± 0.06)%, 50μmol / L group (0.48 ± 0.06%) and 100μmol / L cyclopentamine group (0.92 ± 0.11)% (P <0.05). The result of Western blot showed that 10μmol / L omepamine group (0.78 ± 0.06)%, 50μmol / L omepminamine group (0.58 ± 0.05)% and 100μmol (0.40 ± 0.04)% Gli-1 protein expression was significantly lower than that of the blank control group (0.94 ± 0.11)% (P <0.05). Conclusion Hedgehog signaling pathway inhibitor Wangboming can inhibit glioma U87 cell proliferation and induce apoptosis of U87 cells, which may be related to the down-regulation of GLi-1 expression.